GSM7802520: control_2; Homo sapiens; RNA-Seq
Total RNA of triplet replicates were isolated from control and stably expressing GFP-TFEB Hela 3G cells using QIAschredder (Qiagen, 79645) and RNeasy extraction kit (Qiagen, 74104) according to the manufacturer's instruction. RNA integrity was analyzed with the RNA 6000 Nano Chip on an Agilent 2100 Bioanalyzer (Agilent Technologies) and mRNA was extracted using the NEBNext Poly(A) mRNA Magnetic Isolation module (New England Biolabs). RNA-Seq libraries were generated using the NEXTFLEX Rapid Directional qRNA-Seq Kit (Bioo Scientific) according to the manufacturer's recommendations. All samples' concentration was measured with a Qubit 2.0 Fluorometer (ThermoFisher Sci), and fragment lengths distribution of the final libraries were analyzed with the DNA High Sensitivity Chip on an Agilent 2100 Bioanalyzer (Agilent Technologies). Libraries were multiplexed sequenced on the NextSeq500 (Illumina) with 1x75bp, with 20 to 22 Mio reads per sample.